Therapeutic bandage

ABSTRACT

A therapeutic bandage includes a bandage matrix and an array of microneedles extending from the bandage matrix. Each of the microneedles includes a first layer that encapsulates a first immunomodulatory compound and a second layer that encapsulates a second immunomodulatory compound. The array of microneedles is configured to guide foreign agents affected by the first immunomodulatory compound, the second immunomodulatory compound, or the first and second immunomodulatory compounds from one or more skin layers of a user to the bandage matrix such that the bandage matrix absorbs and captures the foreign agents.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 62/987,494, filed Mar. 10, 2020, the entire contents of which isincorporated by reference herein.

BACKGROUND

Chronic non-healing skin wounds and skin and soft tissue infections(SSTIs) such as those caused by methicillin resistant Staphylococcusaureus (MRSA) and other bacteria are easily acquired in a variety ofsettings (e.g., daycare facilities, college dorm rooms, long-term carefacilities, and hospitals or other healthcare establishments). SSTIs aretypically treated topically, with or without high-dose antibiotictherapy. If improperly treated, these infections can penetrate deeperlayers of skin, necessitating more aggressive surgical remedies toremove puss and necrotic tissue and to properly irrigate the wound.Moreover, the infection may enter blood vessels, allowing it to becomeestablished in distant tissues as well as potentially life-threateningsepsis. There are approximately 120,000 hospitalizations and 20,000deaths per year attributable to MRSA infections, for example, but also11.6 million ambulatory care visits per year for SSTIs, many of whichare the result of chronic infections. Current treatments for theseinfections are costly and frequently ineffective. That is, currentlyavailable bandages do very little by way of immune modulation or thesequestration of toxins and microorganisms. Moreover, current topicaltreatment approaches for treating MRSA, and other SSTIs, are frequentlyineffective for the following reasons: (i) poor drug delivery to dermaltissue due to the barrier function of the stratum corneum; (ii) thedevelopment of antibiotic-resistant bacterial strains including MRSA andVRSA (vancomycin-resistant Staphylococcus aureus), and (iii) asub-optimal approach for manipulating the immunological microenvironmentwithin the dermal tissue. Also, the current standard of care oftenincludes minor surgery on the infection (incision and drainage) followedby high-dose oral antibiotic therapy. This approach is invasive andexpensive and can result in off-target complications includingdisruption of the intestinal microbiome resulting in life threatening C.difficile infections. Further, the overuse of antibiotics is drivingmany microorganisms to develop antibiotic resistance, a phenomenon thatthe World Health Organization characterizes as ‘one of the biggestthreats to global health, food security, and development today. Finally,the known treatments are often not effective in achieving completebacterial clearance, so infections often recur.

SUMMARY

In one embodiment, a therapeutic bandage is provided including a bandagematrix and a plurality of microneedles extending from the bandagematrix, each of the plurality of microneedles including a first layerthat encapsulates a first immunomodulatory compound and a second layerthat encapsulates a second immunomodulatory compound. The first layer isconfigured to release the first immunomodulatory agent at a first rateand the second layer is configured to release the secondimmunomodulatory agent at a second rate that is slower than the firstrate. The first layer is positioned at a distal end of the second layer,and the second layer defining a channel extending from the distal end tothe bandage matrix. The bandage matrix includes a hydration layer and asequestration layer, the hydration layer is configured to absorb aforeign agent removed from a skin infection and the sequestration layeris configured to bind to the foreign agent removed. The bandage matrixincludes a cellulose layer that is configured to bond to a biofilmresulting from the skin infection.

In another embodiments, a therapeutic bandage includes a bandage matrixand an array of microneedles extending from the bandage matrix. Each ofthe microneedles includes a first layer that encapsulates a firstimmunomodulatory compound and a second layer that encapsulates a secondimmunomodulatory compound. The array of microneedles is configured toguide foreign agents affected by the first immunomodulatory compound,the second immunomodulatory compound, or the first and secondimmunomodulatory compounds from one or more skin layers of a user to thebandage matrix such that the bandage matrix absorbs and captures theforeign agents.

In another embodiment, a therapeutic bandage includes a bandage matrixand at least one biodegradable microneedle extending from the bandagematrix. The at least one microneedle includes a base including a firstend that is coupled to the bandage matrix, a second end opposite thefirst end, and a channel extending therethrough from the first end tothe second end, and a tip coupled to the second end of the base. The tipis formed from a first material that encapsulates a firstimmunomodulatory compound and the base is formed from a second materialthat encapsulates a second immunomodulatory compound. The first materialis configured to dissolve at a first rate and the second material isconfigured to dissolve at a second rate that is less than the firstrate. The first immunomodulatory agent and the second immunomodulatoryagent establish a chemotactic gradient within one or more skin layers.The channel is configured to guide foreign agents affected by the firstimmunomodulatory compound, the second immunomodulatory compound, or thefirst and second immunomodulatory compounds from the one or more skinlayers of a user to the bandage matrix such that the bandage matrixabsorbs and captures the foreign agents.

In another embodiment, a method of treating a skin infection or skincondition is provided including administering, via a first layer of amicroneedle, a first immunomodulatory compound beneath the skin,administering, via a second layer of the microneedle, a secondimmunomodulatory compound beneath the skin, and draining phagocyticcells effected by the first immunomodulatory compound and the secondimmunomodulatory compound through a channel in the microneedle.

In another embodiment, a method of treating a skin infection or skincondition in humans and animals includes administering, via a firstlayer of a microneedle, a first immunomodulatory compound to a firstlayer of the skin or a biofilm layer, administering, via a second layerof the microneedle, a second immunomodulatory compound to a second layerof skin or the biofilm layer, and absorbing, by a bandage matrix,foreign agent affected by the first immunomodulatory compound, thesecond immunomodulatory compound, or the first and secondimmunomodulatory compounds. The second layer of the skin may be the sameor different than the first layer of the skin.

Other aspects of the invention will become apparent by consideration ofthe detailed description and accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

FIG. 1A illustrates a schematic view of a therapeutic bandage accordingto one embodiment.

FIG. 1B illustrates a schematic of a portion of the therapeutic bandageof FIG. 1A.

FIG. 1C illustrates another schematic view of the therapeutic bandage ofFIG. 1A.

FIG. 1D illustrates a schematic of a portion of the therapeutic bandageof FIG. 1A.

FIG. 2A illustrates a portion of the therapeutic bandage of FIG. 1A andinteraction with underlying tissue.

FIG. 2B illustrates the portion of the therapeutic bandage of FIG. 1Aand interaction with underlying tissue.

FIG. 2C illustrates the portion of the therapeutic bandage of FIG. 1Aand interaction with underlying tissue.

FIG. 3A illustrates a portion of the therapeutic bandage of FIG. 1A.

FIG. 3B illustrates another view the portion of the therapeutic bandageof FIG. 3A.

FIG. 3C illustrates another view the portion of the therapeutic bandageof FIG. 3A.

FIG. 4A illustrates a first portion of the therapeutic bandage of FIG.1A.

FIG. 4B illustrates the first portion of FIG. 4A coupled to a secondportion of the therapeutic bandage of FIG. 1A.

FIG. 4C illustrates a transverse section of explanted human skinfollowing a therapeutic bandage application.

FIG. 4D illustrates a longitudinal section of explanted human skinshowing spacing of microneedle deposition in the dermal layer followingthe therapeutic bandage application.

FIG. 4E illustrates explanted human skin showing contents released byfollowing the therapeutic bandage application.

FIG. 5 illustrates the epifluorescence microscopy results after contentsfrom the therapeutic bandage are released into the skin layers ofexplanted skin.

DETAILED DESCRIPTION

Before any embodiments of the invention are explained in detail, it isto be understood that the invention is not limited in its application tothe details of construction and the arrangement of components set forthin the following description or illustrated in the following drawings.The invention is capable of other embodiments and of being practiced orof being carried out in various ways.

FIGS. 1A and 1B illustrate a therapeutic bandage 10 including a first orsubcutaneous portion 14 that extends through outer layer (e.g.,epidermis) of the skin 18 of a user and is positioned under the skin 18.The therapeutic bandage 10 also includes a second portion 22 that ispositioned on or above the skin 18. The first portion 14 includes aplurality of dual-layer microneedles 26 (only one needle of theplurality of needles is shown in FIG. 1A), and the second portion 22includes a bandage matrix 30. The plurality of dual-layer microneedles26 are arranged in an array of microneedles 26 (e.g., an array with adensity of about 1,200 microneedles per square inch of the bandagematrix 30). The plurality of microneedles 26 are at least temporarilycoupled to the bandage matrix 30. The bandage matrix 30 is positionableor coupleable to a surface of the skin 18 such that the plurality ofmicroneedles 26 penetrate the skin 18. That is, the microneedles 26penetrate through the epidermis layer of the skin 18 (and the biofilm,if present) such that a portion becomes lodged in the dermis layer ofthe skin 18. The dermis layer is vascular.

With further reference to FIGS. 1A, 1B, 4A, and 4B, each of theplurality of microneedles 26 include a tip 40 (e.g., first needle layer)and a base 44 (e.g., second needle layer). The tip 40 is formed from afirst polymeric material and encloses (e.g., suspends, encapsulates) afirst immunomodulatory compound, a biological agent, an antimicrobialagent, or a combination of a immunomodulatory compound, a biologicalagent, and an antimicrobial agent. In the illustrated embodiment, thetip 40 is substantially conical, although the tip 40 may be othersuitable shapes in other or alternative embodiments. The first polymericmaterial dissolves at a first rate. The first polymeric material may be,for example, a mixture of polyvinylpyrrolidone (PVP) and polyvinylalcohol (PVA), although the first polymeric material may be any suitablematerial in other or alternative embodiments.

The base 44 includes a first (e.g., distal) end coupled to the tip 40, asecond end coupled to the bandage matrix 30, and a channel 50 (e.g.,microchannel or aperture) extending therethrough (also see FIG. 4A)between the first end and the second end. In the illustrated embodiment,the base 44 is substantially cylindrical, but in other or additionalembodiments, the base 44 may be any suitable shape. The base 44 isformed from a second polymeric material. The second polymeric materialmay be, for example, poly(lactic-co-glycolic acid) (PLGA), although thesecond polymeric material may be any suitable material in other oralternative embodiments. The base 44 encloses (e.g., suspends,encapsulates) a second immunomodulatory compound. Like the tip 40, thebase 44 may further enclose one or more biological agents, antimicrobialagents or both. The second polymeric material dissolves at a second ratethat is slower than the first rate.

The microneedles 26 are non-toxic and biodegradable. In the illustratedembodiment, the tip 40 has a first length, and the base 44 has a secondlength that is equal to the first length. In the illustrated embodiment,the first length and the second length are 600 microns. In other oralternative embodiments, may have any suitable lengths. For example, inother embodiments, either or both of the first length or the secondlength may range from 100 microns to 600 microns. Moreover, in someembodiments, the total length of the microneedles 26 (e.g., the sum ofthe first length and the second length) may range from 200 microns to1200 microns. Accordingly, and as discussed in greater detail below, themicroneedles 26 are configured to deliver different biologically activecompounds to different tissue depths thereby instigating a specificresponse for a specific type of pathogen or other foreign agent (forexample, and without limitation, neoplasms, liver spots, poison ivy,poison oak, poison sumac, or tattoo ink). In some embodiments, thebiologically active compounds may be instigate a specific response toother types of skin conditions, such as hereditary skin disorders (e.g.,vitiligo), auto-immune disorders (e.g., lupus, scleroderma), and/orage-related degeneration of the skin (e.g., discoloration and/orwrinkling of skin). Additionally, in other or alternative embodiments,the microneedles 26 may include additional layers, which may be formedfrom a polymeric material and may enclose (e.g., suspend, encapsulate)additional immunomodulatory compounds, anti-microbial compounds,biologically active compounds, or a combination thereof. The sequentialdelivery of biologically active materials in close proximity to oneanother (via the co-localization of the tips 40 of the microneedles 26and the base of the 44 of the microneedles 26) facilitates a two-phasemovement or a three-phase movement of leukocytes into the tissue asdescribed below by virtue of establishing chemokine gradients. Thisproximity augments egress of bacteria-laden leukocytes out of thetissue, via the microchannel 50 and into the bandage matrix.

In the illustrated embodiment, each of the plurality of microneedles 26is configured to match or closely match the biological process of theinfectious agent of a MRSA skin infection. In other or additionalembodiments, the microneedles may be configured to match or closelymatch the biological processes of the infectious agents of other typesof infections or other types of foreign agents. In the example of MRSA,MRSA evades host defenses in part by secreting many virulence factors,which disrupt neutrophil function. Neutrophils are recognized as the keyhost effector cell population for phagocytosing and killing MRSA. MRSAcounteracts neutrophil function with an arsenal of its own, whichincludes neutrophil-killing toxins such as the Panton-Valentineleucocidin (PVL), alpha-toxin, phenol-soluble modulins, among others. Inthe initial stages of MRSA infection, the bacterium is only mildlypathogenic, growing in small planktonic microcolonies. Upon sensing abacterial community, these microcolonies secrete quorum-sensing signalsvia a Agr two-component regulatory system and switch to a hyper-virulentform while initiating the formation of impenetrable biofilms. One goalof the therapeutic bandage 10 is to prevent this switch using a newclass of ‘virulence inhibiting’ antibiotics, as discussed below.Neutrophils traffic into inflamed tissues following a chemotacticgradient in response to bacterial products (e.g.,N-formylmethionyl-leucyl-phenylalanine (fMLF)), or chemokines (e.g.,interleukin-8 (IL-8)) released by leukocytes and other cells, often inresponse to tissue damage.

Although only described in the context of a single microneedle 26, thefollowing process applies to each of the microneedles 26 of thetherapeutic bandage 10. As shown in FIG. 2A, after application of thebandage 10 to the site of interest, the polymeric material of the tip 40begins to dissolve at the first rate, which is about 15 minutes to 30minutes. As the tip 40 dissolves, the first immunomodulatory compound,the biological agent, and the antimicrobial agent encapsulated in thefirst polymeric material are released into the tissue beneath the skin18. As the tip 40 dissolves, neutrophils migrate out of local capillarybeds and into the tissue. In the illustrated embodiment, the firstimmunomodulatory compound is a chemokine (or combination of chemokines),which influences the migration of white blood cells (e.g., macrophagesand neutrophils) from the blood stream into the infected area. In theillustrated embodiment, the first immunomodulatory compound is IL-8having a concentration ranging from 1 nM to 10 nM and fMLF having aconcentration of ranching 200 nM to 1,000 nM, which may induce robustneutrophil infiltration and priming/activation. In other embodiments,any of the known classes of chemokines or combinations of classes ofchemokines may be used. Known, classes of chemokines include, but arenot limited to, lipids (e.g., PGE2, platelet activating factor (PAF)),N-formylated peptides (e.g., bacterial, mitochondrial, or other FPR1, 2and 3 agonists, such as fMLF, fMMYALF), eicosinoids (e.g., lipoxin A4)and other small molecules (e.g. pepducins, host-derived peptides,complement anaphylotoxins (C5a), or small proteins peptides), orproteins (e.g., CXCL8 and/or CXCL2), small molecules includingleukotrienes (e.g., LTB4), cytokines (e.g., interleukin-8 (IL-8),IL-17A/IL-23), methylated BSA and/or any other suitable compound.

The biological agent helps the body fight the bacteria or infectionagent (e.g., MRSA) and facilitate wound disinfection by “opsinizing”bacteria to enhance phagocytosis, and/or by attracting elements of thecomplement system. Opsinization is the process by which microorganismsare ‘made tasty’ to phagocytes upon coating of their outer surface withantibodies or complement components. Moreover, the biological agent canalso neutralize bacterial toxins. The biological agent may include anorganism-specific monoclonal antibody such as anti-Gmd (or othersuitable MRSA-specific antibody), anti-MecA (PBP2a), anti-alpha toxin,and/or any suitable organism-specific monoclonal antibody (e.g.,monoclonal antibody). In one example, monoclonal antibodies target aMRSA surface protein known as glucoseaminidase (gmd), which dramaticallyimproves phagocytosis of planktonic MRSA as well as MRSA growing inmegaclusters. This so-called ‘opsonophagocytic activity’ is initiatedwhen the Fc portion of the monoclonal antibody is recognized byFc-receptors (or CD14) expressed on the plasma membrane of neutrophils,triggering bacterial internalization.

The antimicrobial agents help disable the microorganism to facilitatetheir uptake by the naturally phagocytic cells. In the illustratedembodiment, the antimicrobial agents may include vancomycin, daptomycin,a combination of vancomycin and daptomycin, sitafloxicin, and/or anysuitable antimicrobial agent or antibiotic (e.g., apicidin, savirin,ambuic acid, or any other member of this class of antibiotics, which mayfurther include hydroxyketones, oxacillin, peptide-conjugated lockednucleic acids, tetrapeptide derivatives, ω-hydroxyemodin, or acombination thereof). Other or additional antimicrobial agents may beused instead of or in addition to other or additional embodiments. Thecells effected by the first immunomodulatory compound, the biologicalcompound, and the antimicrobial agent (e.g., phagocytic cells) begin todisinfect the tissue beneath the skin. It should be noted that theantibiotic dose is very low in comparison to current practice(administered orally or IV), and will remain localized to the skin(predominantly) thereby avoiding many of the consequences associatedwith high-dose antibiotic therapy, especially on the gut microbiome. Theuse of very low doses of locally administered antibiotics, and the useof ‘virulence inhibitors’ prevents the development of antibioticresistance since these agents provide no growth/survival advantage tobacteria (unlike traditional bactericidal/bacteriostatic antibiotics).For example, the inhibition (via a virulence inhibitor antibiotic) ofthe Agr two-component quorum-sensing regulatory system prevents biofilmproduction and the expression of nearly 200 downstream virulence genes,many of which inhibit neutrophil function. Moreover, the inhibition ofquorum sensing signals helps maintain MRSA in a planktonic state, makingthe bacterium much more susceptible to phagocytosis.

With reference to FIGS. 1B, 1D, 2B, and 2C, once the tip 40 dissolves,the base 44 remains such that the channel 50 is accessible. Then thepolymeric material of the base 44 begins to dissolve at the second rate,which is about 24 hours to about 72 hours. As the base 44 dissolves, thesecond immunomodulatory compound begins to release from the base 44, andcells effected by the contents of the tip 40 migrate towards the channel50. More specifically, neutrophils (and/or other white blood cells,e.g., macrophages) released as the tip 40 dissolves acquire thepathogenic bacterial (e.g., infectious agent, foreign agent) within thewound and begin their migration towards the channel 50 due to therelease of the second immunomodulatory compound. Because the channel 50has the highest concentration of immunomodulatory compounds within andaround an opening 54 (FIG. 1B) thereof, the phagocytic cells move towardand through the opening 54 (FIG. 2C). Accordingly, the phagocytic cells(with their recently-acquired bacterial, infectious agent or foreignagent payload) travel through the channel 50 due to capillary fluidmovement, exit through the skin 18, and enter the bandage matrix 30 thatis resting or adhered to the surface of the skin 18. In the illustratedembodiment, the second immunomodulatory compound is substantiallysimilar to that of the first immunomodulatory compound. Accordingly, thesecond immunomodulatory compound is a combination of chemokines.Specifically, in the illustrated embodiment, the second immunomodulatorycompound is IL-8 having a concentration ranging from 1 nM to 10 nM andfMLF having a concentration ranging from 200 nM to 1,000 nM. The secondimmunomodulatory compounds may be different than the firstimmunomodulatory compounds in other embodiments. Also, other oradditional immunomodulatory compounds may be used instead of or inaddition to the chemokines. Moreover, the biological agents, theantimicrobial agents or both discussed above may be used. If used, thebiological agents, the antimicrobial agents, or both used in the base 44may be the same or different than the biological agents, theantimicrobial agents, or both used in the tip 40.

With reference to FIGS. 1A, 1B, 2, and 3 , the phagocytic cells (whichoften includes necrotic tissue, infectious agents, foreign agents,bacterial toxins, cellular debris and autolysis fluids) that movethrough the channel 50 of the base 40 of the microneedle 26 are absorbedand captured by the bandage matrix 30, which will be discussed ingreater detail below.

The bandage matrix 30 includes a transparent film layer 60 (e.g., abarrier film layer formed from a polyurethane membrane), a first bandagelayer 62 (e.g., hydration layer), a second bandage layer 64 (e.g.,sequestration layer), and a wound contact (e.g., cellulose) layer 70.The layers may be coupled by adhesive (e.g., acrylic adhesive) or othersuitable methods.

The hydration layer 62 defines a hydrodynamic gradient based on fluidcapillary action using adsorbent hydrogel materials (e.g., calciumalginate) to hasten fluid efflux from the wound into the bandage matrix30. In some embodiments, a third immunomodulatory compound may beincluded in the hydration layer to further attract phagocytic cells intodeeper layers of the bandage matrix 30. In some embodiments, thehydration layer 62 may be formed from hydrogel impregnated with thethird immunomodulatory compound (e.g., chemokine-impregnated), which isdiscussed in detail below. In the illustrated embodiment, the thirdimmunomodulatory compound is substantially similar to that of the firstand second immunomodulatory compounds. Accordingly, in the illustratedembodiment, the third immunomodulatory compound is IL-8 having aconcentration ranging from 1 nM to 10 nM and fMLF having a concentrationranging from 200 nM to 1,000 nM. The third immunomodulatory compound maybe different than the first and second immunomodulatory compounds inother embodiments. Moreover, biological agents, antimicrobial agents, orboth discussed above may be also be included in the hydration layerused. If used, the biological agents, the antimicrobial agents, or bothused in the hydration layer 62 may be the same or different than thebiological agents, the antimicrobial agents, or both used in the tip 40and the base 44.

The sequestration layer 64, which is shown in FIGS. 3A-3B, may bepositioned adjacent to the hydration layer 62. In the illustratedembodiment, the sequestration layer 64 includes a base layer 90 that iscoupled to or coated with a capture layer 94 having one or moreimmobilized antibodies 98. Also, a ligand or dye 102 is bound to the oneor more of the antibodies 98 as part of a detection/saturation reportersystem. In the illustrated embodiment, the base layer 90 is 6%cross-linked agarose bound to protein A/G and the capture layer 94includes monoclonal antibodies configured to attract or otherwisetightly adhere to the respective infectious agents or foreign agents(e.g., MRSA in this embodiment). The sequestration layer 64 sequestersthe infectious agents or foreign agents and associated toxins. The dye102 bound to the antibodies is released when the infectious agent orforeign agent is present. The dye 102 that is released diffuses radiallyinto the hydration layer 62 becoming visible through the transparentfilm, thereby alerting the patient or healthcare worker that thetherapeutic bandage 10 has reached its capacity and should be replaced.

In one specific embodiment, as wound exudate migrates into thetherapeutic bandage 10, it will pass through the cellulose layer 70(discussed below) and will encounter the base layer (e.g., the diffusematrix of 6% beaded agarose covalently modified with Protein A or G),which binds to antibody Fc regions with high avidity. In thisembodiment, the antibodies are monoclonal antibodies to Gmd, which willbind to and sequester any free bacteria which enter the bandage matrix30. During manufacture, approximately 10% of the antibody binding siteswill be occupied with recombinant Gmd-conjugated to 400 nm blue latexbeads 102. As the capacity of the bandage 10 for MRSA binding isapproached, the blue latex beads 102 will gradually be released,allowing them to diffuse into the hydrogel 62 where they will becomevisible to the naked eye through the clear, medical grade polyurethanebarrier film. As noted above, this modified lateral flow immunoassaywill report bandage saturation to the health care professional,triggering bandage removal and replacement.

The cellulose layer 70 provides a mechanism for micro-debridement as thecarbohydrate layer created by the cellulose intermingle with acarbohydrate layer of the infectious agent (e.g., the exopolysaccharideof the MRSA present in the biofilm at the surface of the wound) orforeign agent. That is, a biofilm developed by the wound (which oftenimpedes proper wound healing) may grow into the cellulose layer,creating permanent points of attachment. Once the therapeutic bandage 10is removed, the biofilm will remain associated (integral) with thecellulose layer 70, providing a mechanism for pain-free removal,obviating the need for surgical debridement. In addition, the celluloselayer 70 can serve as a reservoir for additional chemokines andanti-microbial agents should lab tests indicate such a need.

Many superficial skin wounds become chronically infected, which leads tothe breakdown of the epidermis due to constant contact with bacterialenzymes and host derived toxic factors (e.g. reactive oxygen/nitrogen,proteolytic enzymes etc.). This can lead to the formation of ‘chronic,non-healing wounds’ requiring frequent surgical debridement procedureswhich are painful and expensive. The bacteria which colonize thesewounds, including MRSA, in response to sensing quorum factor signalsexit planktonic growth patterns and form biofilms which are nearlyimpregnable by small molecules due to the production of anexopolysaccharide protective outer shell. Accordingly, in someembodiments, the cellulose layer 70 be made of a cellulose((C6H10O5)_(x)) (25 micron pore size) into which the exopolysaccharidewill attach and intertwine with the cellulose matrix. Upon removal, thebiofilm will remain attached to the cellulose layer 70, providing amechanism for micro-debridement without surgical intervention. In otherembodiment, a second approach to prevent biofilm production may be inthe impregnation of the hydration layer (which may include 12.5% calciumalginate hydrogel) with anti-virulence therapeutic agents such asapicidin or other suitable antimicrobial agent or antibiotic (discussedabove), which, by virtue of inhibiting the agr system will also preventthe transition into biofilm production. Alginate, for example, is anaturally occurring anionic and hydrophilic polysaccharide comprised ofcrosslinked (1-4)-linked β-d-mannuronic acid (M) and α-1-guluronic acid(G) monomers. Impregnation of calcium alginate with apicidin and thethird immunomodulatory compound may be accomplished using supercriticalCO₂, among other processes. Once the hydrogel becomes water-saturated(approximately 30-fold swelling), directional movement of water willcease, and fluid flow within the bandage 10 will become static. Thus,the remaining activity will be based on chemotaxis and drug diffusion.The impregnation of the third immunomodulatory compound may allowbacteria-laden (e.g., infectious agent-laden, foreign agent-laden,MRSA-laden) neutrophils to move several millimeters into the bandagematrix, providing a unique approach for disinfecting the wound. Takentogether, these innovations may also speed healing by virtue ofpreventing ‘collateral damage’ from virulence factors normally releasedby the bacterium, and by preventing the formation of a surface biofilm.

As multiple applications of the therapeutic bandage may be needed, thepenetration points of the microneedles will differ, allowing punctatehealing over the entire wound bed. Thus, different portions of the woundmay be at different stages in the disinfection/wound-healing process.Over time (and with repeated applications), the wound will besufficiently disinfected to allow normal wound healing to proceed in anunhindered fashion.

FIG. 4A illustrates a microneedle base 44 showing the hollow channel 50.FIG. 4B illustrates the microneedle base 44 coupled to therapid-dissolving tips 40, which include tattoo ink. In one embodiment,the therapeutic bandage 10 may be producing the microneedles 26 and thencoupling the microneedles 26 to the bandage matrix 30. In someembodiments, the microneedles 26 may be integrally formed with a portionof the bandage matrix 30. The microneedles are produced by forming thebase 44, with the microchannel 50, and loading the base 22 with thesecond immunomodulatory compound. The tip 40 is then produced and loadedwith the first immunomodulatory compound. The tip 40 is then coupled tothe base 44 to seal the microchannel 26. To test the mechanical strengthof the microneedles 26, the tips 40 were loaded with insoluble tattooink (FIG. 4B). The microneedles 26 were then applied to cultured humanskin explants using a 1 Newton application force. The tips 40 weresufficiently rigid to penetrate the epidermis (FIG. 4C) and deliver thetattoo ink in close proximity to dermal blood vessels (‘BV’ in FIG. 4D).That is, FIG. 4C shows a transverse section of explanted human skinfollowing bandage application, indicating the depth of penetration andpayload deposition of insoluble tattoo ink into the dermal layer. Morespecifically, FIG. 4C shows the depth of tissue penetration anddeposition of the ink after an insertion force of 1N. FIG. 4D shows theink in the dermal layer adjacent to blood vessels in explained humanskin. FIG. 4E illustrates a longitudinal section of explanted human skinshowing spacing of microneedle deposition in the dermal layer. Thekinetics of drug release were then measured using epifluorescencemicroscopy and two fluorescent dyes. BODIPY-vancomycin (green) wasloaded in the tips 40, while sulforhodamine B was loaded into the base44. As anticipated, the tips 40 dissolved within 30 minutes anddelivered the BODIPY-vancomycin into the dermal layer of skin, with adiffusion radius of approximately 440 microns to 600 microns (FIG. 5 atA). By one hour after bandage application, the burst release ofDODIPY-vancomycin (green) from the tip 40 had diffused and was no longerdetectable. Conversely, the red fluorescence was released much moreslowly (after about one hour) from the microneedle base 44, withapproximately equal diffusion radius (FIG. 5 at D). Moreover, as shownat FIG. 5 at B, the sulforhodamine B dye was barely visible after only30 minutes.

The therapeutic bandage 10 discussed herein is suitable for use inhealthcare settings including hospitals, outpatient clinics and nursinghomes, as well as for over-the-counter applications andprescription-based applications for skin infections and conditions. Alsoin addition to being appropriate for any bacterial, fungal or viral skininfection, the therapeutic bandage 10 discussed herein may be suitablefor the foreign agents of other skin conditions, such as poisonivy/oak/sumac, minor burns, tattoo removal, bio-threat agents, acne,contact sensitivities and allergic conditions including eczema, insectstings/bites, diabetic foot ulcers, pressure ulcers, venous ulcers, etc.Additionally, as noted above, the therapeutic bandage 10 discussedherein may be suitable other types of skin conditions, such ashereditary skin disorders (e.g., vitiligo), auto-immune disorders (e.g.,lupus, scleroderma), and/or age-related degeneration of the skin (e.g.,discoloration and/or wrinkling of skin). Pets and other companionanimals (e.g. horses and some farm animals) also suffer fromdermatological skin conditions including allergic eczema, MRSAinfections and others. One embodiment of the current invention could beused to treat such animals by formulating the device withspecies-specific therapeutic agents in keeping with accepted veterinarypractices.

The therapeutic bandage 10 discussed herein provides a highly targetedlocal therapy and promotes more rapid healing. It relies on the preciseadministration of immunomodulatory compounds to orchestrate the movementof leukocytes and other cell populations known or suspected to beassociated with wound disinfection and healing, both temporally andspatially. That is, the therapeutic bandage 10 i) recruits high numbersof neutrophils (and/or other types of white blood cells, e.g.,macrophages), the key type of white blood cell needed to clear theinfection, into the infected tissue; (ii) manipulates the immunologicalenvironment within the dermal tissue to prevent neutrophil (and/or othertypes of white blood cell, e.g., macrophage) killing and maximize theirability to engulf and destroy the bacteria; and (iii) provides amechanism for interstitial fluid movement, which facilitates the removalof pus, detritus, neutrophils (and/or other types of white blood cells,e.g., macrophages), bacterial toxins and bacteria to exit the wound andbecome entrapped in the bandage matrix 30. The plurality of microneedles26 (i) deliver therapeutic agents (e.g., the first and secondimmunomodulatory compounds) into the dermal layer of infected skin topromote neutrophil (and/or other types of white blood cell, e.g.,macrophage) movement into the wound plus additional agents to ensuretheir survival and maximizing their function; (ii) have a bi-directionalchannel allowing fluid communication between the bandage matrix 30 andthe infected tissues to facilitate the egress from the wound, and (iii)a four-layer bandage matrix which entraps (e.g., absorbs and captures)wound exudate paired with a reporter system to alert healthcare workerswhen the bandage becomes saturated. The elements referenced aboveprovide a ‘three-phased chemokine’ approach, which establishes atemporal and spatial gradient of chemo-attractants. The first wave ofimmunomodulatory compounds from the tip 40 of the microneedle 26facilitates the movement of neutrophils (and/or other types of whiteblood cells, e.g., macrophages) first from blood into the wound,followed hours later by the second wave of immunomodulatory compoundsemanating from the base 44 of the microneedle 26. The co-localization ofthe tips 40 of the microneedles 26 and the channel 50 is key to properfunctioning helps funnel bacteria-laden (e.g., infectious agent-laden,foreign agent-laden) neutrophils (and/or other types of white bloodcells, e.g., macrophages) toward the channel 50, where interstitialfluid movement will sweep them out of wound and into the bandage matrix30. Once net fluid movement slows to a stop, the third wave ofimmunomodulatory compounds emanating from the water saturated hydrationlayer 62 will attract neutrophils (and/or other types of white bloodcells, e.g., macrophages) into deeper layers of the bandage matrix 30.The immunomodulatory compounds, the biological agents, and theantimicrobial agents maximize neutrophil infiltration, prevent theirinactivation, and hasten their movement out of the wound followinguptake of infectious agents (or other foreign agents) therebyfacilitating rapid wound disinfection and healing and preventing thespread of infectious organisms to visceral organs and to otherindividuals.

Moreover, the therapeutic bandage 10 discussed herein is minimallyinvasive and prevents dissemination of infectious organisms to visceraltissues, thereby mitigating a potentially life-threatening condition.For example, the microneedles 26 are small enough to avoid touchingnerve endings, significantly reducing pain while penetrating the stratumcorneum to enable effective drug delivery into the highly vascularizeddermal layer of skin. In some instances, the therapeutic bandage can beself-administered by a patient, offering a high degree of patientcompliance at a significantly reduced cost.

Various features and advantages of the invention are set forth in thefollowing claims.

1-20. (canceled)
 21. A therapeutic bandage comprising: a bandage matrix;and an array of microneedles extending from the bandage matrix, each ofthe microneedles including a first layer that encapsulates a firsttherapeutic agent and a second layer that encapsulates a secondtherapeutic agent, wherein the array of microneedles is configured todeliver at least one of the first therapeutic agent or the secondtherapeutic agent from the bandage matrix into the skin of a patient.22. The therapeutic bandage of claim 21, wherein the bandage matrixcomprises a first bandage layer disposed on a film layer, a secondbandage layer disposed on the first bandage layer, and a cellulose layerdisposed on the second bandage layer, the array of microneedlesprotruding from the cellulose layer.
 23. The therapeutic bandage ofclaim 21, wherein the first layer is configured to release the firsttherapeutic agent at a first rate, and the second layer is configured torelease the second therapeutic agent at a second rate that is less thanthe first rate.
 24. The therapeutic bandage of claim 21, wherein thefirst layer defines a first length and the second layer defines a secondlength, the first layer being configured to release the firsttherapeutic agent at a first tissue depth, the second layer beingconfigured to release the second therapeutic agent at a second tissuedepth that is different than the first tissue depth.
 25. The therapeuticbandage of claim 21, wherein the first layer is formed from a mixture ofpolyvinylpyrrolidone (PVP) and polyvinyl alcohol (PVA) and the secondlayer is formed from poly(lactic-co-glycolic acid) (PLGA).
 26. Thetherapeutic bandage of claim 21, wherein at least one of the firsttherapeutic agent and the second therapeutic agent comprise at least oneof an immunomodulatory compound, a biological agent or an antimicrobialagent.
 27. The therapeutic bandage of claim 26, wherein theimmunomodulatory compound comprises at least one of: chemokines, lipids,N-formylated peptides, eicosinoids, leukotrienes, cytokines, ormethylated BSA.
 28. The therapeutic bandage of claim 26, wherein thebiological agent comprises at least one of an organism specificmonoclonal antibody, anti-MecA anti-alpha toxin, or an organism-specificmonoclonal antibody.
 29. The therapeutic bandage of claim 26, whereinthe antimicrobial agent comprises at least one of vancomycin,daptomycin, sitafloxicin, apicidin, savarin, ambuic acid,hydroxyketones, oxacillin, peptide-conjugated locked nucleic acids,tetrapeptide derivatives, ω-hydroxyemodin, or a combination thereof. 30.A method for fabricating a therapeutic bandage, the method comprising:forming a bandage matrix; and disposing an array of microneedlesextending from the bandage matrix, each of the microneedles including afirst layer that encapsulates a first therapeutic agent and a secondlayer that encapsulates a second therapeutic agent, wherein the array ofmicroneedles is configured to deliver at least one of the first agent orthe second agent from the bandage matrix into the skin of a patient. 31.The method of claim 30, wherein the forming comprises: disposing a firstbandage layer on a film layer; disposing a second bandage layer on anopposing side of the first bandage layer; disposing a cellulose layer onan opposing side of the second bandage layer; and, configuring themicroneedles to protrude from the cellulose layer.
 32. The method ofclaim 30, further comprising disposing within at least some of themicroneedles at least one of an immunomodulatory compound, a biologicalagent or an antimicrobial agent.
 33. A therapeutic bandage comprising: abandage matrix comprising a first bandage layer disposed on a filmlayer, a second bandage layer disposed on the first bandage layer and acellulose layer disposed on the second bandage layer; and an array ofmicroneedles extending from the cellulose layer of the bandage matrix,each of the microneedles including a first layer that encapsulates afirst therapeutic agent and a second layer that encapsulates a secondtherapeutic agent, wherein the array of microneedles is configured todeliver at least one of the first therapeutic agent or the secondtherapeutic agent from the bandage matrix into the skin of a patient.34. The therapeutic bandage of claim 33, wherein at least one of thefirst therapeutic agent and the second therapeutic agent comprise atleast one of an immunomodulatory compound, a biological agent or anantimicrobial agent.